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課程名稱 |
重組DNA技術
Recombinant Dna Technology |
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開課學期 |
110-2 |
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授課對象 |
生物資源暨農學院 園藝暨景觀學系 |
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授課教師 |
杜宜殷 |
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課號 |
HORT5031 |
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課程識別碼 |
628 U1640 |
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班次 |
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學分 |
3.0 |
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全/半年 |
半年 |
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必/選修 |
選修 |
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上課時間 |
星期二A,B(18:25~20:10)星期三10,A,B(17:30~20:10) |
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上課地點 |
轉溫101轉溫101 |
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備註 |
初選不開放。作物群必選。本系學生優先。請先向授課教師登記。
總人數上限：6人 |
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課程簡介影片 |
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核心能力關聯 |
本課程尚未建立核心能力關連 |
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課程大綱
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為確保您我的權利,請尊重智慧財產權及不得非法影印
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課程概述 |
1. Inoculation and culture of bacteria in solid and liquid media
2. Plating of bacteriophage
3. Extraction of bacteriophage DNA
4. Phenol extraction and quantitation of bacteriophage DNA
5. Restriction digestion of bacteriophage DNA
6. Electrophoresis and recovery of digested bacteriophage DNA from agarose gel
7. Quantitation of recovered DNA and ligation
8. Bacterial transformation
9. Determination of efficiency of transformation
10. Minipreparation of plasmid DNA |
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課程目標 |
This course attempts to provide the opportunity for senior graduate students to learn recombinant DNA techniques including principles, methods, equipments, and chemicals. The experimental materials are E. coli and bacteriophage. |
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課程要求 |
tudents who have taken courses related to molecular biology or biotechnology |
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預期每週課後學習時數 |
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Office Hours |
另約時間 |
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指定閱讀 |
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參考書目 |
1. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (2004) Current Protocols in Molecular Biology. by John Wiley & Sons, Inc. Chichester, UK.
2. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (2003) Short Protocols in Molecular Biology, 5th ed. John Wiley & Sons, Inc. New York.
3. Brown TA (2010) Gene Cloing and DNA analysis – An introduction. 6th ed. Blackwell Science Ltd, London.
4. Clark DP (2005) Molecular Biology - Understanding the Genetic Revolution. Elsevier Academic Press, USA.
5. Glick BR, Pasternak JJ, Patten CL (2010) Molecular Biotechnology: Principles and Applications of Recombinant DNA. American Society for Microbiology Press, Washington, DC.
6. Hackett PB, Fuchs JA, Messing JW (1984) An Introduction to Recombinant DNA Techniques. The Benjamin/Cummings Publishing Company, Inc.
7. Hardin C, Edwards J, Presutti D, Miller W, Robertson D (2001) Cloning, Gene Expression, and Protein Purification. Oxford University Press, Inc.
8. Micklos DA, Freyer GA (2003) DNA Science: A First Course. 2nd ed. Cold Spring Harbor Laboratory Press, New York.
9. Miesfeld RL (1999) Applied Molecular Genetics. Wiley-Liss, Inc.
10. Rapley R (2000) The Nucleic Acid Protocols Handbook. Humana Press Inc.
11. Rodriguez RL, Tait RC (1983) Recombinant DNA Techniques: an introduction. Addison-Wesley Publishing Company.
12. Sambrook J, David W, Sambrook J (2001) Molecular Cloning: a laboratory manual, 3nd ed, Vol. 1 and 3. Cold Spring Harbor Laboratory Press, New York. |
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評量方式
(僅供參考) |
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週次 |
日期 |
單元主題 |
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第1週 |
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課程介紹 |
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第2週 |
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大腸桿菌固體與液體培養基之配製 |
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第3週 |
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大腸桿菌之固體與液體培養 |
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第4週 |
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噬菌體培養基之配製 |
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第5週 |
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噬菌體之灑佈培養 |
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第6週 |
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噬菌體之純化 |
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第7週 |
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溫書假 |
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第8週 |
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噬菌體DNA之抽取 |
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第9週 |
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噬菌體DNA去RNA處理及定量 |
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第10週 |
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噬菌體DNA限制酶切 |
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第11週 |
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瓊脂洋菜凝膠電泳分析及膠體內DNA片段之回收 |
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第12週 |
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回收DNA片段之估量與連結反應 |
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第13週 |
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勝任細胞之製備 |
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第14週 |
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轉型質體至大腸桿菌 |
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第15週 |
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轉型效率之估算、藍白篩選之觀察及接種 |
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第16週 |
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重組質體DNA之小量製備 + 期末考 |
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第17週 |
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重組質體DNA之酶切及電泳檢測 |
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第18週 |
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期末考 |
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